Synthesis and antitumor evaluation of 5-(benzo[: D] [1,3]dioxol-5-ylmethyl)-4-(tert -butyl)- N -arylthiazol-2-amines

Article abstract of DOI:10.1039/c6md00234j

A series of novel N-aryl-5-(benzo[d][1,3]dioxol-5-ylmethyl)-4-(tert-butyl)thiazol-2-amines (C1-C31) were synthesized and evaluated for their antitumor activities against HeLa, A549 and MCF-7 cell lines. Some tested compounds showed potent growth inhibition properties with IC₅₀ values generally below 5 μM against the three human cancer cells lines. Compound C27 showed potent activities against HeLa and A549 cell lines with IC₅₀ values of 2.07 ± 0.88 μM and 3.52 ± 0.49 μM, respectively. Compound C7 (IC₅₀ = 2.06 ± 0.09 μM) was the most active compound against A549 cell line, while compound C16 (IC₅₀ = 2.55 ± 0.34 μM) showed the best inhibitory activity against the MCF-7 cell line. The preliminary mechanism of the inhibitory effect was investigated via further experiments, such as morphological analysis by dual AO/EB staining and Hoechst 33342 staining, and cell apoptosis and cycle assessment by FACS analysis. The results illustrated that compound C27 could induce apoptosis and cause both S-phase and G2/M-phase arrests in HeLa cell line. Therefore, compound C27 could be developed as a potential antitumor agent.

Full text of DOI:10.1039/c6md00234j

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DOI: 10.1039/C6MD00234J
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RESEARCH ARTICLE
Synthesis and antitumor evaluation of 5‐(benzo[d][1,3]dioxol‐5‐
ylmethyl)‐4‐(tert‐butyl)‐N‐arylthiazol‐2‐amines
Z. L. Wua, Y. L. Fanga, Y. T. Tanga, M. W Xiaoa, J. Yea, G. X. Lia, and A. X. Hu*a
Received 00th January 20xx,
Accepted 00th January 20xx
A series of novel N‐aryl‐5‐(benzo[d][1,3]dioxol‐5‐ylmethyl)‐4‐(tert‐butyl)thiazol‐2‐amines (C1‐C31) were synthesized and
evaluated for their antitumor activities against HeLa, A549 and MCF‐7 cell lines. Some tested compounds showed potent
growth inhibition properties with IC50 values generally below 5 μM against the three human cancer cells lines. Compound
C27 showed potent activities againt HeLa and A549 cell lines with IC50 of 2.07 ± 0.88 μM and 3.52 ± 0.49 μM respectively.
Compound C7 (IC50 = 2.06 ± 0.09 μM) was the most active compound against A549 cell lines, while compound C16 (IC50
=
DOI: 10.1039/x0xx00000x
2.55 ± 0.34 μM) showed the best inhibitory activity against MCF‐7 cell line. The preliminary mechanism of the inhibitory
effect was investigated via further experiments, such as morphological analysis by dual AO/EB staining and Hoechst 33342
staining, cell apoptosis and cycle assessment by FACS analysis. The results illustrated that the antitumor activity of
compound C27 might be associated with its induced S‐phase and G2/M‐phase arrests and apoptosis in HeLa cell line, and
could be developed as a potential antitumor agent.
www.rsc.org/
positive chronic myelogenous leukemia in chronic phase (CP-
CML) in 2010. Dabrafenib,32-34 another anticancer agent
containing thiazole moiety, was approved in 2013 as a single
1. Introduction
Natural products have been highly pursued for screening
anticancer agents in last decades and it is clear that natural
products will continue to be a major source of new structural
leads, and effective drug development depends on
multidisciplinary collaborations.1-4 Recently, the identification
and application of biologically relevant fragments derived from
biologically validated natural products has been recognized as a
powerful strategy in searching new drugs.5 Benzodioxole exists
in many bioactive plant and marine natural products such as
V600E
agent for treatment of BRAF
mutation-positive
unresectable or metastatic melanoma and for use in
combination with Trametinib to treat patients with unresectable
or metastatic melanoma with a BRAF V600E or V600K mutation by
FDA. In recent years, thiazole moiety was always introduced
into novel molecules for developing new anticancer agents.35-37
N
-Aryl thiazole derivatives were found to possess potent
antitumor activities.38-40 We have been involved in the area of
thiazole derivatives for the past few years and various of
thiazole derivatives with antitumor activities were synthesized
successfully.41 In continuation of the previous studies, and
according to molecular hybridization,42, 43 benzodioxole from
bioactive natural products, thiazole from Dasatinib and
Dabrafenib were introduced into a series of new molecules as
anticancer agents (Fig. 1). All the synthesized compounds were
tested for their cytotoxicities against HeLa, A549 and MCF-7
cell lines. The preliminary mechanism of the inhibitory effect
was also investigated via a series of further experiments.
Liriodenine,6,7
Nitidine
chloride,8,9
Sanguinarine,10-12
Berberine13, Podophyllotoxin14,
and its analogues,16,
15
17
Harringtonine18, Homoharringtonine,19-21 and Trabectedin,22,23
which are found to possess potent antitumor activity.
Thiazole is an active moiety in many clinical drugs such as
third generation of broad-spectrum cephalosporin antibiotic
Cefdinir,24 dopamine D2 antagonist Pramipexole,25 non-
steroidal
anti-inflammatory
drugs
Meloxicam26
and
Sudoxicam,27 and antimicrobial agent Abafungin.28 Dasatinib29-
31
is an anticancer agent containing thiazole moiety, that
acquired the approval for the treatment of adult patients with
CP-CML resistant or intolerant to prior therapy in 2006 and
newly diagnosed adult patients with Philadelphia chromosome-
2. Results and discussion
2.1 Chemistry
For the synthesis of target compounds C1
intermediate 1-(benzo[ ][1,3]dioxol-5-yl)-2-bromo-4,4-
dimethylpentan-3-one was synthesized starting from
benzodioxole according to the procedures reported in our
—C31, the key
a. College of Chemistry and Chemical Engineering, Hunan University, No.1 Lushan
Road(s), Changsha, 410082, China. Email: wuzhilin@hnu.edu.cn,
b12110055@hnu.edu.cn, tangyuting14@hnu.edu.cn, mwxiao@hnu.edu.cn,
yejiao@hnu.edu.cn, liguoxi@hnu.edu.cn, axhu@hnu.edu.cn
†Electronic Supplementary Information (ESI) available: Additional details on the
experimental part of the synthesis and biological assays. Selected spectra are
included. See DOI: 10.1039/x0xx00000x
d
(A)
previous study.44
N
-aryl thioureas (B1
—
B31) were synthesized
This journal is © The Royal Society of Chemistry 2016
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N-arylthiazol-2-amines (CVi1ew—ArCtic3le1O)nliinne
by reating benzoyl chloride and ammonium thiocyanate with ylmethyl)-4-(tert-butyl)-
different arylamines, followed by hydrolysis in the presence of yields ranging from 55% to 88%. All the synthesized
aqueous ammonia. The reactions of compound with B1
compounds were characterized by 1H and 13C NMR
B31 were carried out under reflux in ethanol or acetone for spectroscopy and elemental analyses. Compound C31 was
about 8 h to afford the desired 5-(benzo[ ][1,3]dioxol-5- characterized by X-ray single crystal diffraction.
DOI: 10.1039/C6MD00234J
A
—
d
Fig. 1 Design strategy of the target compounds. Compound C27 is the most active compounds against HeLa cell line.
Plane 2 and Plane 3 is 8.087 (42) °, almost parallel. The crystal data
and structure refinement details are listed in Table 1.
Scheme 1 Synthesis of N‐aryl‐5‐(benzo[d][1,3]dioxol‐5‐ylmethyl)‐4‐
(tert‐butyl)thiazol‐2‐amines (C1—C31). Reagents and conditions: (a) i)
POCl3/DMF, 85 °C, 24 h; ii) pinacolone, NaOH, EtOH, 40 °C, 6 h; iii) H2,
Pd‐C, r.t., 4 h; iv) CuBr2, CHCl3/EtOAc, reflux, 4h; (b) i) PhCOCl, NH4SCN,
CH3COCH3, reflux, 2‐5 h; ii) NH3∙H2O, CH3OH, reflux, 2‐4 h; (c) EtOH or
CH3COCH3, reflux, 5‐8 h.
Fig. 2 Crystal Structure of compound C31
.
2.2 Crystal structure
2.3 Biological activity assay
The X-ray crystallographic study showed that compound C31
belonged to the monoclinic crystal system, space group I12/a1.
Selected bond lengths and bond angles are listed in Table S1. As
shown in Fig. 2, the molecule structure is formed of three planes,
including the benzodioxole (Plane 1), thiazole ring (Plane 2), and
quinoline ring (Plane 3). The dihedral angles between Plane 1 and
Plane 2 is 88.366 (55)°, almost perpendicular, while the one between
2.3.1 In vitro antitumor potency
The cytotoxicities of compounds C1
—C31 against three human
tumor cell lines including HeLa, A549 and MCF-7 cells were
tested by the typical 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-
2-H-tetrazolium bromide (MTT) assay. The IC50 values of
compounds C1
—C31 against the three tested tumor cell lines
were shown in Table 2.
2 | Med. Chem. Commun, 2016, 00, 1‐6
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Table 1. Crystal data and structure refinement details for compound C31.
The results indicated that most of the tested compounds
View Article Online
DOI: 10.1039/C6MD00234J
showed better activity against the three tumor cell lines
Empirical formula
Formula weight
Temperature
Wavelength
Crystal system
Space group
C25H25N3O2S
431.54
150(2) K
0.71073 Å
Monoclinic
I12/a1
a = 19.2951(9) Å α = 90°.
b = 13.6381(4) Å β = 119.209(5)°
c = 19.3808(8) Å γ = 90°
4451.5(3) Å3
comparing with Fluorouracil (5-FU) and Taxol, two known
marketable anticancer drugs. Compounds C16, C24, C27 and
C30 showed the best inhibitory activities against HeLa cell
lines with IC50 values below 5 μM. Compounds C7 and C27
were the most potent inhibitors against A549 cell lines with
IC50 values of 2.06 ± 0.09 and 3.52 ± 0.49 μM respectively. For
MCF-7 cell line, much more compounds showed strongly
potent inhibitory activities with IC50 values below 5 μM. The
structure-activity-relationship revealed that the characteristics
Unit cell dimensions
Volume
Z
Density (calculated)
Absorption coefficient
F(000)
Crystal size
Theta range for data collection
Index ranges
Reflections collected
Independent reflections
Completeness to theta = 26.00° 99.8 %
8
1.288 Mg/m3
of
compounds. For compounds with
locations of substituents on -phenyl has a pronounced effect
on the bioactivities of the tested compounds. In general,
compounds with monosubstituted -phenyls were more
N
-Aryl could significantly influence the bioactivities of the
0.172 mm‐1
N
-phenyls, the amounts and
1824
0.41 × 0.38 × 0.37 mm3
2.99 to 26.00°
‐23 ≤ h ≤ 23, ‐16 ≤ k ≤ 10, ‐23 ≤ l≤ 23
12031
N
N
effective than polysubstituted, while 4-substituted compounds
showed better activities than 2-substituted ones. Compound
4371 [R(int) = 0.0251]
C30 with
N-pyridin-2-yl showed good activities against all of
Absorption correction
Semi‐empirical from equivalents
Max. and min. transmission
Refinement method
0.9390 and 0.9327
the three tumor cell lines, with IC50 values of 4.79 ± 0.86, 5.13
± 0.62, and 4.71 ± 0.47 μM, at the highest level of all the tested
compounds. Compared with pyridin-2-yl and certain substituted
Full‐matrix least‐squares on F2
4371 / 14 / 297
Data / restraints / parameters
Goodness‐of‐fit on F2
1.046
phenyls, naphthanlen-1-yl (C29) and quinolin-8-yl (C31
)
Final R indices [I > 2sigma(I)]
R indices (all data)
Largest diff. peak and hole
R1 = 0.0430，wR2 = 0.1074
R1 = 0.0538，wR2 = 0.1164
0.411 and ‐0.319 e.Å‐3
tended to reduce the antitumor activities of the compounds.
Table 2 Cytotoxicities of compounds
C
on three human tumor cell lines.
IC50 (μM)a
Compound Ar
HeLa
A549
MCF‐7
C1
C2
C3
C4
C5
C6
C7
C8
C6H5
2‐CH3C6H4
3‐CH3C6H4
2‐FC6H4
4‐FC6H4
2‐ClC6H4
3‐ClC6H4
4‐ClC6H4
2‐BrC6H4
3‐BrC6H4
4‐BrC6H4
3‐CF3C6H4
11.95 ± 0.51
24.78 ± 0.79
18.79 ± 1.74
18.32 ± 0.91
25.26 ± 1.75
18.89 ± 3.92
8.27 ± 0.89
10.98 ± 1.91
51.95 ± 0.21
6.43 ± 3.02
13.31 ± 1.89
12.96 ± 0.88
10.36 ± 0.52
11.71 ± 0.62
20.42 ± 1.21
18.74 ± 5.98
12.72 ± 0.16
7.57 ± 0.22
5.81 ± 0.27
6.84 ± 0.21
2.06 ± 0.09
9.19 ± 1.03
26.10 ± 1.58
5.00 ± 0.94
10.05 ± 1.78
15.80 ± 1.94
9.22 ± 1.64
10.12 ± 0.05
11.60 ± 1.14
19.10 ± 2.09
4.91 ± 0.03
4.49 ± 0.49
14.98 ± 1.54
8.99 ± 0.97
3.00 ± 0.25
3.48 ± 0.12
39.85 ± 5.29
4.66 ± 0.25
4.71 ± 0.81
3.32 ± 0.31
3.54 ± 1.90
9.36 ± 1.68
> 200
2.55 ± 0.34
5.95 ± 0.37
18.65 ± 0.77
20.14 ± 5.07
20.37 ± 0.48
19.45 ± 3.30
11.23 ± 0.11
17.72 ± 1.63
8.66 ± 0.38
C9
C10
C11
C12
C13
C14
C15
C16
C17
C18
C19
C20
C21
C22
C23
C24
C25
C26
C27
C28
C29
C30
C31
5‐FU
Taxol
4‐CF3C6H4
4‐OCH3C6H4
2‐NO2C6H4
4‐NO2C6H4
4‐CO2EtC6H4
2,4‐(CH3)2C6H3
2,6‐(CH3)2C6H3
3,4‐(CH3)2C6H3
2,3‐Cl2C6H3
3,4‐Cl2C6H3
3,5‐Cl2C6H3
4‐F‐3‐ClC6H3
4‐NO2‐2‐FC6H3
4‐NO2‐2‐ClC6H3
3,5‐(CF3)2C6H3
2,6‐Br2‐4‐OCF3C6H2
naphthalen‐1‐yl
pyridin‐2‐yl
194.11 ± 44.34 82.80 ± 1.70
4.15 ± 0.01
47.53 ± 27.47
18.90 ± 0.54
83.50 ± 6.15
20.71 ± 5.06
34.61 ± 7.28
7.83 ± 2.17
5.80 ± 0.59
4.60 ± 0.52
19.29 ± 2.13
51.70 ± 11.52
2.07 ± 0.88
53.26 ± 12.37
14.05 ± 0.09
4.79 ± 0.86
8.26 ± 0.18
8.51 ± 4.30
11.35 ± 0.52
15.94 ± 2.07
12.15 ± 2.72
14.24 ± 1.49
11.54 ± 0.37
17.03 ± 1.82
16.30 ± 1.68
67.24 ± 11.72 33.33 ± 1.55
80.88 ± 3.32
3.52 ± 0.49
76.55 ± 1.52
9.03 ± 0.96
102.74 ± 4.25 102.86 ± 2.17
55.19 ± 4.50
5.13 ± 0.62
55.50 ± 0.07
26.00 ± 6.00
21.02 ± 3.00
22.30 ± 0.63
4.71 ± 0.47
> 200
79.30 ± 5.11
13.99 ± 1.01
2‐methylquinolin‐8‐yl 65.07 ± 0.92
56.50 ± 3.40
19.31 ± 3.40
a IC50 are presented as the mean (IC50 ± SEM) of three independent experiments performed in duplicate.
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DOI: 10.1039/C6MD00234J
staining and obvious change of cell morphology, such as
crescentic and ring-shaped cell mass. The apoptosis tended to
be more significant as the concentration increased. In general,
compound C27 caused more obvious apoptosis than compound
C24. The Hoechst 33342 staining analysis suggested that
compounds C24 and C27 could induce apoptosis in HeLa cell
lines, which was consistent with the results of dual AO/EB
staining assay.
2.3.2 Morphological analysis by dual AO/EB staining
Acridine orange (AO) can pass through intact cell membranes
and make cell glow green fluorescence. Ethidium bromide (EB)
can only pass through damaged cell membranes, and glows
orange fluorescence after combined to DNA.
Compounds C24 and C27
, two of the most active
compounds against HeLa cell line, were selected to determine
whether their inhibitory activities were related to the apoptosis-
inducing effect, morphological assay was then performed using
dual AO/EB staining method. The effect of compounds C24
and C27 on the morphological changes of HeLa cells was
shown in Fig. 3. After 24 h of incubation, HeLa cells treated
with C24 and C27 exhibited typical apoptosis features, such as
obvious chromatin condensation and slight nuclear
fragmentation. With the increase of the concentration of
compounds C24 and C27, more apoptotic cells with typical
nuclear fragmentation (orange) appeared. In addition, in the two
tested representatives, compound C27 was more effective in
inducing apoptosis of HeLa cells. The results of AO/EB
staining revealed that compounds C24 and C27 could induce
the apoptosis of HeLa cells. The dosedependent relationship of
the apoptotic cells was consistent with the previous inhibitory
activity assay.
Fig. 4 Images of Hoechst 33342 stained HeLa cells after 24 h of incubation.
The negative control group was treated with DMSO. The experimental
groups were treated with compounds C24 and C27 at 5 μM and 10 μM
respectively.
2.3.4 Apoptosis assessment by FACS analysis
Apoptosis, one of the essential features of cells, plays a
significant role in embryo development, tissue repair and
maintaining homeostasis. Phosphatidylserine (PS) is actively
held facing the cytosolic (inner) side of the cell membrane by
the enzyme flippase. However, when a cell undergoes apoptosis,
PS is no longer restricted to the cytosolic side by flippase and
flips to the extracellular (outer) surface of the cell. Annexin V
is a calphobindin that can specifically bind with the PS flipped
out to the external surface of the cell membrane. In the bivariate
flow cytometry scatter plot, normal cells are in Q4 quadrant
(FITC-/PI-), early apoptotic cells are in Q3 quadrant
(FITC+/PI-), late apoptotic cells are in Q2 quadrant
(FITC+/PI+), while dead cells are in Q1 quadrant (FITC-/PI+).
Fig. 3 Images of AO/EB stained HeLa cells after 24 h of incubation. The
negative control group was treated with DMSO. The experimental groups
were treated with compounds C24 and C27 at 5 μM and 10 μM respectively.
Compounds C24 and C27 were selected as two
representatives to analyze the apoptosis ratios quantitatively by
using fluorescein isothiocyanate (FITC)-conjugated Annexin V
and propidium iodide (PI) double staining method on a flow
cytometer. As shown in Fig. 5 and 6, after a 24-hour treatment
of C24 and C27, the percentage of early apoptotic cells
increased from 2.36% (control) to 11.7% (C24, 5 μM), 19.5%
2.3.3 Morphological analysis by Hoechst 33342 staining
Hoechst 33342 is a blue fluorescent dyes which can pass
through cell membranes. It’s easier for Hoechst 33342 to enter
into apoptotic cells than normal ones.
In order to observe the changes of nuclei morphology of
HeLa cells after the treatment of compounds C24 and C27
,
Hoechst 33342 staining was performed and the results were
shown in Fig. 4. Slightly blue and homogeneous cells were
observed in control group. While in groups treated with
compounds C24 and C27, cells showed bright blue nuclear
(
C24, 10 μM) and 19.2% (C27, 5 μM), 62.7% (C27, 10 μM)
respectively, while the percentages of late apoptotic cells
maintained at a level below 5%. The results revealed that
compound C24 and C27 could induce apoptosis of HeLa cells
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obviously in a dosedependent manner. Summarizing the
aforementioned results in dual AO/EB staining, Hoechst 33342
staining, and cell apoptosis assessment by FACS analysis, it
suggested that compound C24 and C27 inhibited HeLa cells’
growth by inducing apoptosis.
View Article Online
DOI: 10.1039/C6MD00234J
Fig. 7 Cell cycle analysis of HeLa cell lines after 24 h of incubation. The
negative control group was treated with DMSO. The experimental groups
were treated with compounds C24 and C27 at 5 μM and 10 μM respectively.
Fig. 5 Apoptosis of HeLa cells after 24 h of incubation. The negative control
group was treated with DMSO. The experimental groups were treated with
compounds C24 and C27 at 5 μM and 10 μM respectively.
Fig. 8 Quantitative analysis of the effect of different concentrations of
compounds C24 and C27 on cell cycle in HeLa cells compared with control
group.
Conclusions
A series of novel
N-aryl-5-(benzo[d][1,3]dioxol-5-ylmethyl)-4-
Fig. 6 Quantitative analysis of the effect of different concentrations of
compounds C24 and C27 on inducing apoptosis in HeLa cells compared with
control group.
(
tert-butyl)thiazol-2-amines were synthesized and evaluated
their cytotoxicities against HeLa, A549 and MCF-7 human
cancer cell lines. Quite a few of the tested compounds showed
potent growth inhibition properties with IC50 values generally
below 5 μM against the three cancer cell lines. Compounds C27
C7 and C16 showed the best inhibitory activities against HeLa,
A549 and MCF-7 cell lines respectively, with IC50 = 2.07 ±
0.88 μM, 2.06 ± 0.09 μM and 2.55 ± 0.34 μM. To investigate
the mechanism of the inhibitory effect, compounds C24 and
C27 were selected as representatives to perform a series of
biological assays such as morphological analysis by dual
AO/EB staining and Hoechst 33342 staining, cell apoptosis and
cycle assessment by FACS analysis in HeLa cell line. The
results illustrated that compounds C24 and C27 could induce
cell apoptosis and mainly S-phase arrest in HeLa cell lines in a
dose-dependent manner. Moreover, compound C27 could also
cause G2/M-phase arrest in HeLa cell line.
,
2.3.5 Cell cycle assessment by FACS analysis
Cell cycle analysis of HeLa cells treated with compounds C24
and C27 were performed to investigate whether they exerted
the inhibitory effects by inducing cell cycle arrest. As shown in
Fig. 7 and 8, significant S-phase arrests were observed in HeLa
cells treated with compounds C24 and C27 at 5 and 10 μM for
24 h. In the groups treated with compound C24, the proportion
of HeLa cells in S phase increased from 23.69% (control) to
28.69% (5 μM) and 36.63% (10 μM) respectively. In the groups
treated with compound C27, the proportion of HeLa cells in S
phase increased from 23.69% (control) to 27.10% (5 μM) and
35.93% (10 μM) respectively and the proportion of HeLa cells
in G2/M phase increased from 9.89% (control) to 16.03% (5
μM) and 17.93% (10 μM) respectively. The result suggested
that compound C24 mainly induced cell cycle S-phase arrest
while compound C27 induced both S-phase and G2/M-phase
arrests in cell cycles.
Acknowledgements
The authors would like to thank Professor Jiannan Xiang
(College of Chemistry and Chemical Engineering, Hunan
University) and Professor Kemin Wang (State Key Laboratory
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Stroiakovski, M. Lichinitser, R. Dummer, F. Grange, L. Mortier, V. Chiarion‐
of Chemo/Biosensing and Chemometrics, Hunan University)
for their helps in biological assays.
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Sileni, K. Drucis, I. Krajsova, A. Hauschild, P. Lorigan, P. Wolter, G. V. Long, K.
DOI: 10.1039/C6MD00234J
Flaherty, P. Nathan, A. Ribas, A.‐M. Martin, P. Sun, W. Crist, J. Legos, S. D.
Rubin, S. M. Little and D. Schadendorf, New Engl. J. Med., 2014, 372, 30‐39.
A. R. Ali, E. R. El‐Bendary, M. A. Ghaly and I. A. Shehata, Eur. J. Med.Chem.,
2014, 75, 492‐500.
K. M. Dawood and S. M. Gomha, J. Heterocyclic Chem., 2015, 52, 1400‐
1405.
G. Turan‐Zitouni, M. D. Altintop, A. Ozdemir, Z. A. Kaplancikli, G. A. Ciftci
and H. E. Temel, Eur. J. Med. Chem., 2016, 107, 288‐294.
C. B. Andersen, Y. Wan, J. W. Chang, B. Riggs, C. Lee, Y. Liu, F. Sessa, F. Villa,
N. Kwiatkowski, M. Suzuki, L. Nallan, R. Heald, A. Musacchio and N. S. Gray,
ACS Chem. Biol., 2008, 3, 180‐192.
S. Carradori, D. Rotili, C. De Monte, A. Lenoci, M. D'Ascenzio, V. Rodriguez,
P. Filetici, M. Miceli, A. Nebbioso, L. Altucci, D. Secci and A. Mai, Eur. J. Med.
Chem., 2014, 80, 569‐578.
Z. M. Nofal, E. A. Soliman, S. S. Abd El‐Karim, M. I. El‐Zahar, A. M. Srour, S.
Sethumadhavan and T. J. Maher, J. Heterocyclic Chem., 2014, 51, 1797‐
1806.
J.‐m. Peng, W. Li, K. Shen, S.‐f. Huo, J. Ye and A.‐x. Hu, Chem. J. Chinese U.,
2013, 34, 1646‐1652.
B. S. P. Claudio Viegas‐Junior, B. S. P. Amanda Danuello, B. S. P. Vanderlan
da Silva Bolzani, B. S. P. Eliezer J. Barreiro and B. S. P. Carlos Alberto
Manssour Fraga, Curr. Med. Chem., 2007, 14, 1829‐1852.
C. Karthikeyan, V. R. Solomon, H. Lee and P. Trivedi, Biomedicine &
Preventive Nutrition, 2013, 3, 325‐330.
Z. Wu, N. Ding, D. Lin, A. Hu, J. Ye and G. Li, Chem. Res. Chinese U., 2016, 32,
49‐54.
35.
36.
37.
38.
Notes and references
1.
2.
3.
4.
G. M. Cragg and D. J. Newman, BBA‐Gen. Subjects, 2013, 1830, 3670‐3695.
R. Duffy, C. Wade and R. Chang, Drug Discov. Today, 2012, 17, 942‐953.
D. J. Newman and G. M. Cragg, J. Nat. Prod., 2012, 75, 311‐335.
I. Zupko, W. Jaeger, Z. Topcu and C. C. Wu, Biomed. Res. Int., 2015, 2015, ID
242070. DOI: 10.1155/2015/242070.
5.
B. Yu, Y.‐C. Zheng, X.‐J. Shi, P.‐P. Qi and H.‐M. Liu, Anti‐Cancer Agent. Me.,
2016, 16, 1‐10.
39.
40.
6.
H.‐C. Chang, F.‐R. Chang, Y.‐C. Wu and Y.‐H. Lai, Kaohsiung J. Med. Sci., 2004,
20, 365‐371.
7.
T. J. Hsieh, T. Z. Liu, C. L. Chern, D. A. Tsao, F. J. Lu, Y. H. Syu, P. Y. Hsieh, H. S.
Hu, T. T. Chang and C. H. Chen, Food Chem. Toxicol., 2005, 43, 1117‐1126.
Z. Fang, Y. Tang, W. Jiao, Z. Xing, Z. Guo, W. Wang, Z. Xu and Z. Liu, Food
Chem. Toxicol.,, 2014, 66, 210‐216.
8.
41.
42.
9.
M. Sun, N. Zhang, X. Wang, C. Cai, J. Cun, Y. Li, S. Lv and Q. Yang, Tumor.
Biol., 2014, 35, 10201‐10212.
10.
11.
12.
13.
14.
15.
16.
M. K. Break, M. I. Tahir, K. A. Crouse and T. J. Khoo, Bioinorg. Chem. Appl.,
2013, 2013, ID 362513. DOI: 10.1155/2013/362513.
E. L. Leung, X. X. Fan, M. P. Wong, Z. H. Jiang, Z. Q. Liu, X. J. Yao, L. L. Lu, Y. L.
Zhou, L. F. Yau, V. P. Tin and L. Liu, Antioxid. Redox. Sign., 2016, 24, 263‐279.
S. Pallichankandy, A. Rahman, F. Thayyullathil and S. Galadari, Free Radical
Bio. Med., 2015, 89, 708‐720.
L. M. Ortiz, P. Lombardi, M. Tillhon and A. I. Scovassi, Molecules, 2014, 19,
12349‐12367.
A. Roy, M. J. Ernsting, E. Undzys and S.‐D. Li, Biomaterials, 2015, 52, 335‐
346.
43.
44.
R. Yadav, D. Kumar, A. Kumari and S. K. Yadav, J. Microencapsul., 2014, 31,
211‐219.
K. A. L. Marshall, H. S. H. Chan and D. Simpson, Blood, 2014, 124, 5767‐
5767.
17.
18.
K. R. Hande, Eur. J. Cancer, 1998, 34, 1514‐1521.
D. S. Moirangthem, J. C. Borah, S. Laishram, M. C. Kalita and N. C. Talukdar,
Nat. Prod. Res., 2014, 28, 1503‐1506.
19.
S. Philipp, J. Sosna, J. Plenge, H. Kalthoff and D. Adam, Cell. Commun.
Signal., 2015, 13, DOI: 10.1186/s12964‐015‐0103‐0.
M. Wetzler and D. Segal, Curr. Pharm. Design., 2011, 17, 59‐64.
J. Zhang, B. Chen, T. Wu, Q. Wang, L. Zhuang, C. Zhu, N. Fan, W. Qing, Y. Ma
20.
21.
and
X.
Xu,
PLoS
One,
2015,
10,
e0142422.
DOI:
10.1371/journal.pone.0142422 .
22.
23.
M. D'Incalci, N. Badri, C. M. Galmarini and P. Allavena, Brit. J. Cancer, 2014,
111, 646‐650.
S. Avila‐Arroyo, G. S. Nunez, L. F. Garcia‐Fernandez and C. M. Galmarini, J.
Breast Cancer, 2015, 18, 329‐338.
24.
25.
D. R. P. Guay, Clin. Ther., 2002, 24, 473‐489.
K. M. Shannon, J. P. Bennett, J. H. Friedman and t. P. S. Group, Neurology,
1997, 49, 724‐728.
26.
27.
S. Noble and J. A. Balfour, Drugs, 1996, 51, 424‐430; discussion 431‐432.
R. S. Obach, A. S. Kalgutkar, T. F. Ryder and G. S. Walker, Chem. Res. Toxicol.,
2008, 21, 1890‐1899.
28.
29.
C. Borelli, M. Schaller, M. Niewerth, K. Nocker, B. Baasner, D. Berg, R.
Tiemann, K. Tietjen, B. Fugmann, S. Lang‐Fugmann and H. C. Korting,
Chemotherapy, 2008, 54, 245‐259.
E. Jabbour, H. M. Kantarjian, G. Saglio, J. L. Steegmann, N. P. Shah, C. Boqué,
C. Chuah, C. Pavlovsky, J. Mayer, J. Cortes, M. Baccarani, D.‐W. Kim, M. B.
Bradley‐Garelik, H. Mohamed, M. Wildgust and A. Hochhaus, Blood, 2013,
123, 494‐500.
30.
H. M. Kantarjian, N. P. Shah, J. E. Cortes, M. Baccarani, M. B. Agarwal, M. S.
Undurraga, J. Wang, J. J. Kassack Ipiña, D.‐W. Kim, M. Ogura, C. Pavlovsky, C.
Junghanss, J. H. Milone, F. E. Nicolini, T. Robak, J. Van Droogenbroeck, E.
Vellenga, M. B. Bradley‐Garelik, C. Zhu and A. Hochhaus, Blood, 2011, 119,
1123‐1129.
31.
32.
T. Takeda, H. Kanzaki, S. Toyooka, M. Watanabe, T. Ohtsuka, K. Suzawa, S.
Hashida, Y. Maki, H. Yamamoto, J. Soh, H. Asano, K. Tsukuda, S. Miyoshi, Y.
Kitamura and T. Sendo, Cancer Res., 2015, 75, 724.
G. S. Falchook, G. V. Long, R. Kurzrock, K. B. Kim, T. H. Arkenau, M. P. Brown,
O. Hamid, J. R. Infante, M. Millward, A. C. Pavlick, S. J. O’Day, S. C.
Blackman, C. M. Curtis, P. Lebowitz, B. Ma, D. Ouellet and R. F. Kefford,
Lancet, 2012, 379, 1893‐1901.
33.
34.
A. Hauschild, J.‐J. Grob, L. V. Demidov, T. Jouary, R. Gutzmer, M. Millward, P.
Rutkowski, C. U. Blank, W. H. Miller Jr, E. Kaempgen, S. Martín‐Algarra, B.
Karaszewska, C. Mauch, V. Chiarion‐Sileni, A.‐M. Martin, S. Swann, P. Haney,
B. Mirakhur, M. E. Guckert, V. Goodman and P. B. Chapman, Lancet, 2012,
380, 358‐365.
C. Robert, B. Karaszewska, J. Schachter, P. Rutkowski, A. Mackiewicz, D.
6 | Med. Chem. Commun, 2016, 00, 1‐6
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DOI: 10.1039/C6MD00234J
Graphical Abstract
Natural anticancer agents
Dasatinib and Dabrafenib
O
O
Molecular
Hybridization
N
S
CF3
O
O
N
S
N
CF3
H
C27
IC50 = 2.07 0.88 M for HeLa cell line
±
The strategy for designing target compounds as antitumor agents.